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Robotics-Driven Manufacturing of Cartilaginous Microtissues for Skeletal Tissue Engineering Applications.

Isaak Decoene1,2, Gabriele Nasello1,3, Rodrigo Furtado Madeiro de Costa4

  • 1Prometheus Division of Skeletal Tissue Engineering, KU Leuven, Leuven, Belgium.

Stem Cells Translational Medicine
|January 13, 2024
PubMed
Summary

Automated robotics enhance cartilage microtissue manufacturing. This bioprocess integrates robotics for media changes and imaging, ensuring quality for clinical translation of engineered tissues.

Keywords:
AutomationImage processingManufacturinganimal modelsautologousbonechondrogenesisclinical translationprogenitor cellstissue engineering

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Area of Science:

  • Biotechnology
  • Tissue Engineering
  • Regenerative Medicine

Background:

  • Automated manufacturing is crucial for robust production of tissue-engineered products.
  • Robotics offer potential for automating complex bioprocesses in tissue engineering.
  • Current methods for microtissue culture and quality assessment often rely on manual operations.

Purpose of the Study:

  • To develop and validate an automated strategy for culturing and monitoring cartilaginous microtissues.
  • To establish noninvasive quality attributes using automated image analysis.
  • To optimize liquid handling parameters for long-term microtissue differentiation protocols.

Main Methods:

  • Utilized a robotics platform for automated media changes and imaging of microtissues in microwells.
  • Developed an automated image analysis pipeline for microtissue displacement and morphological feature extraction.
  • Employed Design of Experiments (DOE) to optimize liquid handling parameters.
  • Used ovine periosteum-derived cells for chondrogenic differentiation and assessed differentiation markers (COL2A1, RUNX2).
  • Performed histological analysis and ectopic implantation in nude mice.

Main Results:

  • Achieved 96% accuracy and 0.84 Dice coefficient in identifying empty microwells.
  • Identified that repeated media changes and culture duration, not aspiration/dispension speeds, drive microtissue displacement.
  • Confirmed chondrogenic differentiation (COL2A1 expression) without osteogenic specification (RUNX2).
  • Observed increased extracellular matrix and glycosaminoglycan secretion in larger microtissues.
  • Demonstrated successful formation of mineralized tissues and bone from microtissue implants after ectopic implantation.

Conclusions:

  • An integrated automated bioprocess for cartilaginous microtissue culture and manipulation was successfully developed.
  • Automated imaging and analysis provide noninvasive quality control metrics for tissue-engineered products.
  • The developed system shows promise for the future manufacturing of microtissue-based living implants, reducing reliance on manual operations.